本文選題：系統(tǒng)性白念菌感染 + Hsp90　； 參考：《東北師范大學(xué)》2007年碩士論文

【摘要】： 白色念珠菌是一種寄生于健康人體的正常菌群,在機(jī)體的免疫力下降或者正常菌群失調(diào)的情況下,就會(huì)轉(zhuǎn)變成致病菌。免疫抑制劑的應(yīng)用增加了病人對(duì)念珠菌的易感性,使醫(yī)院內(nèi)患者念珠菌感染的發(fā)病率不斷增加。 熱休克蛋白90(Hsp90)是白色念珠菌在侵染機(jī)體過(guò)程中分泌的一種毒性因子,同時(shí),作為念珠菌的一種免疫優(yōu)勢(shì)抗原,可以誘導(dǎo)感染機(jī)體產(chǎn)生特異性抗體。Matthews和本實(shí)驗(yàn)室的工作證明,抗Hsp90的抗體是一種保護(hù)性抗體。 我們課題組已經(jīng)證明,噬菌體展示的表位能夠誘導(dǎo)小鼠產(chǎn)生體液免疫應(yīng)答和細(xì)胞免疫應(yīng)答。在念珠菌感染過(guò)程中,CD4+T細(xì)胞的分化方向是決定機(jī)體對(duì)念珠菌感染抗性或易感性的重要方面。為了進(jìn)一步研究噬菌體作為表位載體,誘導(dǎo)小鼠產(chǎn)生細(xì)胞因子的類(lèi)型,本研究利用展示有C表位的雜合噬菌體、野生噬菌體和與匙孔血藍(lán)蛋白(KLH)偶聯(lián)的化學(xué)合成C表位(KLH-C),免疫C57BL/6J小鼠,再采用定量ELISA方法,檢測(cè)脾細(xì)胞培養(yǎng)上清中分泌細(xì)胞因子的水平。同時(shí)用RT-PCR方法分析了mRNA水平上細(xì)胞因子的轉(zhuǎn)錄,確定了檢測(cè)小鼠體內(nèi)脾中Th1和Th2型細(xì)胞因子的轉(zhuǎn)錄。得到的結(jié)果如下: 1.噬菌體作為疫苗載體可以增強(qiáng)免疫鼠脾細(xì)胞Th1型細(xì)胞因子的轉(zhuǎn)錄。雜合噬菌體、野生噬菌體和KLH-C免疫小鼠體內(nèi)脾中Th1型細(xì)胞因子,IFN-γ、IL-2和IL-12的轉(zhuǎn)錄水平相似,與TE對(duì)照組比較,差異顯著。IL-10在免疫鼠和對(duì)照鼠間的轉(zhuǎn)錄水平相似。 2.噬菌體作為疫苗載體可以誘導(dǎo)免疫鼠的脾細(xì)胞分泌Th1型細(xì)胞因子。雜合噬菌體免疫小鼠的脾細(xì)胞培養(yǎng)上清中,Th1型細(xì)胞因子,IFN-γ和IL-12的蛋白分泌水平與TE對(duì)照組比較,具有顯著性差異,與KLH-C免疫小鼠相似。Th2型細(xì)胞因子,IL-10的分泌水平在免疫組和對(duì)照組之間沒(méi)有差異。 因此,噬菌體作為疫苗載體,能夠通過(guò)增強(qiáng)Th1型細(xì)胞因子的轉(zhuǎn)錄誘導(dǎo)Th1型細(xì)胞因子的產(chǎn)生,與傳統(tǒng)的化學(xué)載體KLH相比,具有相似的作用。而且其制備簡(jiǎn)單、省時(shí)省力的優(yōu)點(diǎn)使噬菌體作為表位載體在疫苗研究中具有廣闊的前景。
[Abstract]:Candida albicans is a normal group of bacteria parasitic on healthy people, which can be transformed into pathogenic bacteria when the immunity of the body decreases or the normal flora is out of balance. The use of immunosuppressants increases the susceptibility of patients to Candida and increases the incidence of Candida infection in hospitals. Heat shock protein 90 (Hsp90) is a toxic factor secreted by Candida albicans during its infection. As an immune dominant antigen of Candida albicans, heat shock protein 90 (Hsp90) can induce infection to produce specific antibodies. Antibody against Hsp90 is a protective antibody. Our team has demonstrated that phage-displayed epitopes can induce humoral and cellular immune responses in mice. In the process of Candida infection, the differentiation direction of CD4 T cells is an important factor in determining the resistance or susceptibility to Candida infection. In order to further study the type of cytokine production induced by phage as epitope carrier, a hybrid phage with C epitope was used in this study. Wild phages and chemically synthesized C epitopes (KLH-C) coupled with keyhole hemocyanin (KLH) were used to immunize C57BL / 6J mice. The levels of cytokines secreted in the supernatant of splenocyte culture were detected by quantitative Elisa. At the same time, the transcription of cytokines at mRNA level was analyzed by RT-PCR, and the transcription of Th1 and Th2 type cytokines in mouse spleen was determined. The results are as follows: 1. Phage as a vaccine vector can enhance the transcription of Th1 cytokines in spleen cells of immunized mice. The transcription levels of IL-2 and IL-12 in spleen of hybrid phage, wild phage and KLH-C immunized mice were similar to those of te control group. The transcriptional level of IL-10 in immunized mice and control mice was similar. 2. 2. Bacteriophage as a vaccine vector can induce Th1 cytokine secretion from spleen cells of immunized mice. The secretory levels of Th1 type cytokines IFN- 緯 and IL-12 in the supernatant of spleen cell culture of hybrid phage immunized mice were significantly different from those of te control group. The secretory level of IL-10 was similar to that of KLH-C immunized mice. There was no difference between the immunized group and the control group. Therefore, phage, as a vaccine vector, can induce Th1 cytokine production by enhancing the transcription of Th1 cytokines, which is similar to that of traditional chemical vector KLH. Moreover, the advantages of simple preparation, time saving and labor saving make bacteriophage as epitope carrier has a broad prospect in vaccine research.
【學(xué)位授予單位】：東北師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類(lèi)號(hào)】：R392

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